LOCUS       sequence-364138-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 Nucleoprotein fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  AUTHORS   May DG, Martin-Sancho L, Anschau V, Liu S, Chrisopulos RJ, Scott KL,
            Halfmann CT, Diaz Pena R, Pratt D, Campos AR, Roux KJ
  TITLE     A BioID-Derived Proximity Interactome for SARS-CoV-2 Proteins.
  JOURNAL   Viruses. 2022 Mar 15;14(3). pii: v14030611. doi: 10.3390/v14030611.
  PUBMED    35337019
REFERENCE   2  (bases 1 to 7066)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Viruses."; 
            date: "2022-03-15"; volume: "14(3). pii: v14030611. doi"; pages: " 
            10.3390/v14030611"
COMMENT     Sequence Label: N-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             31..500
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    565..764
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             965..1381
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1311..1333
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1349..1365
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2682..3377
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             3378..3404
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3419..3439)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3424..3753
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3604..3739
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3763..4362
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3763..3782)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4259..4279
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4583..5052
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5182..5199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5433..5452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6101..6892,6893..6961))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6724..6743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6962..7066)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg atatctgtgg
       61 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
      121 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
      181 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
      241 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
      301 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
      361 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
      421 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
      481 ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg
      541 accaccgacc caccaccggg aggcaagctg gccagcaact tatctgtgtc tgtccgattg
      601 tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg
      661 tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag
      721 acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg
      781 gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc
      841 gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca
      901 gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
      961 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     1021 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     1081 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     1141 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     1201 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1261 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1321 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1381 cggccggatc cgccaccatg agcgataacg gcccccaaaa ccaacggaat gcccctcgga
     1441 taacatttgg tggccctagt gattccacag gaagtaacca aaacggagaa cggtctggag
     1501 cccgctctaa acagaggcga ccacagggat tgccaaataa cacagcgtca tggtttacag
     1561 ccctgactca acatggaaag gaggacttga aatttccacg aggacagggt gtaccaataa
     1621 acacaaacag tagtccagac gatcagatcg ggtattatag aagggccacg agacgcatca
     1681 gaggtggaga tggtaaaatg aaagatctca gtccgcgctg gtacttttat tatttgggaa
     1741 ccggcccaga agcgggcctc ccatatggag ccaataagga cgggattata tgggtggcta
     1801 cagagggagc cctcaatacg ccaaaagatc acatcggaac aagaaaccct gctaataatg
     1861 ccgcaatcgt gctgcagttg cctcagggga cgacgcttcc taaaggcttt tacgcagaag
     1921 gatcacgcgg cggcagccaa gcatcctcaa ggtctagttc cagaagtcga aacagctcca
     1981 gaaactccac accagggtcc agtaggggca caagtccggc gcggatggcg ggcaacggcg
     2041 gagacgccgc actcgctctg ttgctcctgg accggctcaa ccaacttgaa tccaagatga
     2101 gtggtaaggg acagcagcaa caaggtcaaa ccgtaaccaa gaaaagcgct gcagaagctt
     2161 ctaaaaaacc tcgacaaaaa cggaccgcta cgaaggcata taacgttact caagccttcg
     2221 ggagacgagg gccggagcaa acccagggga attttggaga tcaggagctc atccggcaag
     2281 ggacagatta caaacattgg cctcaaatcg ctcagttcgc gccgtccgca agtgcttttt
     2341 tcggaatgtc tcgcatcggt atggaagtaa caccatccgg gacatggctc acttataccg
     2401 gcgccatcaa gctggacgac aaagatccaa atttcaaaga tcaggtgatt ctcttgaata
     2461 aacacataga cgcgtataag acgtttccac ccaccgaacc caagaaggac aaaaagaaaa
     2521 aagctgatga aacacaagcg ttgccccagc gacagaagaa gcagcaaacc gtcaccctcc
     2581 tgccagccgc tgatctcgat gacttctcca agcaacttca acagtccatg agctcagccg
     2641 attctactca ggcgaccggt ggtggaggcg ggtctgaatt cttcaagaac ctgatctggc
     2701 tgaaggaggt ggacagcacc caggagagac tgaaggagtg gaacgtgagc tacggcaccg
     2761 ccctggtggc cgacagacag accaagggca gaggcggcct gggcagaaag tggctgagcc
     2821 aggagggcgg cctgtacttc agcttcctgc tgaaccccaa ggagttcgag aacctgctgc
     2881 agctgcccct ggtgctgggc ctgagcgtga gcgaggccct ggaggagatc accgagatcc
     2941 ccttcagcct gaagtggccc aacgacgtgt acttccagga gaagaaggtg agcggcgtgc
     3001 tgtgcgagct gagcaaggac aagctgatcg tgggcatcgg catcaacgtg aaccagagag
     3061 agatccccga ggagatcaag gacagagcca ccaccctgta cgagatcacc ggcaaggact
     3121 gggacagaaa ggaggtgctg ctgaaggtgc tgaagagaat cagcgagaac ctgaagaagt
     3181 tcaaggagaa gagcttcaag gagttcaagg gcaagatcga gagcaagatg ctgtacctgg
     3241 gcgaggaggt gaagctgctg ggcgagggca agatcaccgg caagctggtg ggcctgagcg
     3301 agaagggcgg cgccctgatc ctgaccgagg agggcatcaa ggagatcctg agcggcgagt
     3361 tcagcctgag aagaagctac ccttacgatg taccggatta cgcatgagtc gaccctgtgg
     3421 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3481 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3541 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3601 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3661 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3721 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc
     3781 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3841 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3901 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3961 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4021 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4081 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4141 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4201 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4261 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4321 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     4381 cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa gattttattt
     4441 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4501 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca
     4561 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     4621 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
     4681 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4741 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     4801 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     4861 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct
     4921 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     4981 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     5041 gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     5101 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca
     6061 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6121 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6181 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6241 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6301 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6361 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6421 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6481 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6541 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6601 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6661 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6721 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6781 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6841 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6901 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7021 acatatttga atgtatttag aaaaataaac aaataggggt tccgcg
//
